The ARC's Molecular/Cellular Biology Laboratory Core will provide a vital role in coordinating the research activities of investigators that have an ARC research component, a pilot study supported by the ARC, or a project supported by the NIAAA that is tied to the theme and objectives of the ARC. ITS mission under the renewal will be to continue to support and maintain a central cell culture facility, to perform analytical procedures in support of research projects, and to expand molecular biology assays and resources deemed to be important to the coordinate efforts of alcohol research as it relates to the overall research objectives of the ARC. The Specific Aims of the Molecular/Cellular Biology Laboratory Core will be: 1. To continue and maintain a cell culture facility and support research activities utilizing cell culture technology. This will support both primary cultures as well as establish cell lines used for cytokine bioassays, and perform bioassays as needed by the ARC participants. 2. To perform specific analytical assays for projects supported by the ARC. These include a) RIA and ELISA procedures for hormones and cytokines; b) ethanol, metabolite, and enzymatic activity assays; c) chromogenic LPS assay. 3. To perform molecular biological assays or procedures as needed by the ARC participants. These include gene expression studies for various rodent and primate cytokines using Northern analysis or semi-quantitative PCR, reverse transcriptase assays for SIV replication detection, and, if required, transfection procedures utilizing viral and non-viral gene transfer methods. To generate and maintain lines of HIV-1 transgenic mice to develop in vivo models of alcohol abuse and HIV pathogenesis in current and future studies. This Aim will include generation and maintenance of lines of transgenic mice that carry HIV-promoter-reporter gene constructs expression one or more of HIV-1 proteins in lymphoid or other tissue. Developmental aspects of the Core will be coordinated through communications with the Intramural Center committee. Priorities will be based on ARC objectives to maximize the research effectiveness of projects supported by the Core. In addition, personnel in the Laboratory Core will provide technical training, coordinate research efforts among projects where appropriate. The overall goal of the Laboratory Core will be to enhance efficiency, quality control, cooperation, and flexibility of research participants in their pursuit of Center objectives.